Imaging systems and methods for tissue differentiation, e.g., for intraoperative visualization

Abstract

Described herein is a multiplex platform that uses ultrasmall nanoparticles (e.g., C dots and C′ dots) to graphically differentiate specific nerves (e.g., sensory nerves vs. motor nerves) for nerve transplants and other surgeries. Also described herein is a multiplex platform that uses ultrasmall nanoparticles (e.g., C dots and C′ dots) to graphically differentiate between different types of lymph nodes and/or lymphatic pathways, e.g., to safely and effectively perform vascularized lymph node transplantation in the treatment of lymphedema. Also described herein is a multiplex platform that uses ultrasmall nanoparticles (e.g., C dots and C′ dots) to graphically differentiate parathyroid tissue.

1 .- 89 . (canceled) 
     
     
         90 . A method comprising:
 administering two or more different probe species, each comprising a spectrally differentiable fluorescent reporter, to a lymphatic system of a human subject with a tumor, wherein a first probe species is administered to a tumor site of the subject, and a second probe species is administered to an extremity of the subject;   directing excitation light into lymph nodes of the subject, thereby exciting the fluorescent reporters having spectrally distinguishable emission wavelengths;   identifying a lymph node that drains the tumor site, and identifying another lymph node that drains the extremity, using a multichannel fluorescence camera system that simultaneously detects the spectrally distinguishable emission wavelengths in real-time; and   displaying in real-time the fluorescent reporters having spectrally distinguishable emission wavelengths to graphically differentiate between the lymph node that drains the extremity and the lymph node that drains the tumor site,   wherein the two or more different probe species each comprise a silica nanoparticle, and   wherein the silica nanoparticle has a diameter less than 10 nm.   
     
     
         91 . The method of  claim 90 , wherein the tumor site comprises a member selected from the group consisting of a breast, a trunk, an abdomen, a pelvis, and a thoracic cavity. 
     
     
         92 . The method of  claim 90 , wherein the extremity comprises a member selected from the group consisting of an upper limb and a lower limb. 
     
     
         93 . The method of  claim 90 , wherein the identifying comprises simultaneously detecting fluorescent light of the spectrally different emission wavelengths, the detected fluorescent light having been emitted by the fluorescent reporters of the respective probe species in the lymph nodes and/or drainage pathways as a result of illumination by the excitation light so as to discriminate between signals received from each probe species. 
     
     
         94 . The method of  claim 90 , wherein the fluorescent reporter of the first probe species having received the excitation light fluoresces at a spectrally distinguishable wavelength compared to the fluorescent reporter of the second probe species having received the excitation light. 
     
     
         95 . The method of  claim 93 , further comprising identifying a lymph node for excision. 
     
     
         96 . The method of  claim 90 , further comprising displaying a map of lymph nodes and/or lymphatic pathways of the lymphatic system, wherein the map graphically differentiates between specific lymph nodes and/or between specific lymph node types. 
     
     
         97 . The method of  claim 96 , wherein the tumor site comprises a member selected from the group consisting of a breast, a trunk, an abdomen, a pelvis, and a thoracic cavity. 
     
     
         98 . The method of  claim 90 , wherein the multichannel fluorescence camera system is a handheld multichannel fluorescence camera system. 
     
     
         99 . A method comprising:
 administering two or more different probe species, each comprising a spectrally differentiable fluorescent reporter, to a lymphatic system of a human subject, wherein a first probe species is administered to a trunk of the subject, and a second probe species is administered to an extremity of the subject;   directing excitation light into lymph nodes of the subject, thereby exciting the fluorescent reporters having spectrally distinguishable emission wavelengths;   identifying a lymph node in the trunk for harvesting in transplantation for lymphedema, and identifying another lymph node that drains the extremity, using a multichannel fluorescence camera system that simultaneously detects the spectrally distinguishable emission wavelengths in real-time; and   displaying in real-time the fluorescent reporters having spectrally distinguishable emission wavelengths to graphically differentiate between the lymph node that drains the extremity and the lymph node for harvesting,   wherein the two or more different probe species each comprise a silica nanoparticle, and   wherein the silica nanoparticle has a diameter less than 10 nm.   
     
     
         100 . The method of  claim 99 , wherein the extremity comprises a member selected from the group consisting of an upper limb and a lower limb. 
     
     
         101 . The method of  claim 99 , wherein the identifying comprises simultaneously detecting fluorescent light of the spectrally different emission wavelengths, the detected fluorescent light having been emitted by the fluorescent reporters of the respective probe species in the lymph nodes and/or drainage pathways as a result of illumination by the excitation light so as to discriminate between signals received from each probe species. 
     
     
         102 . The method of  claim 99 , wherein the fluorescent reporter of the first probe species having received the excitation light fluoresces at a spectrally distinguishable wavelength compared to the fluorescent reporter of the second probe species having received the excitation light. 
     
     
         103 . The method of  claim 99 , further comprising displaying a map of lymph nodes and/or lymphatic pathways of the lymphatic system, wherein the map graphically differentiates between specific lymph nodes and/or between specific lymph node types.